Novel lipid compound, composition and nucleic acid lipid nanoparticle preparation

Abstract

The invention provides an ionizable lipid compound, a composition containing the ionizable lipid compound and application of the ionizable lipid compound. In order to provide more choices for delivery of preparations such as gene drugs, nucleic acid vaccines, small molecule drugs and the like, the invention provides an ionizable lipid compound shown as a general formula # imgabs0 #, or a salt, a prodrug or a stereoisomer of the ionizable lipid compound which can be used by the drugs. The ionizable lipid compound can be used for delivering nucleic acid drugs or small molecule drugs, enriches the types of ionizable lipid compounds, and is of great significance to development and application of nucleic acid preventive and therapeutic agents.

Description

Technical Field

[0001] This application belongs to the field of biomedical technology, and specifically relates to a novel ionizable lipid, which can be used to combine with other lipid components (such as neutral lipids, steroids, and PEG-lipids) to form lipid nanoparticles with the function of in vivo drug delivery. Background Art

[0002] Gene therapy technology is a research hotspot in the field of modern biomedicine. Nucleic acid drugs can be used to prevent cancer, bacterial and viral infections, and treat diseases with genetic etiologies. Due to the characteristics of nucleic acid drugs such as easy degradation and difficulty in entering cells, it is necessary to encapsulate them with the help of a carrier and deliver them to target cells. Therefore, the development of a safe and efficient delivery carrier has become a prerequisite for the clinical application of gene therapy.

[0003] Lipid nanoparticles (LNPs) are currently a research hotspot in the field of non-viral gene carriers. As a class of safe and efficient nucleic acid drug delivery carriers, they have been proven to be able to deliver RNA efficiently to the liver. In 2018, the US FDA approved the world's first LNP-based liver-targeted siRNA drug for the treatment of hereditary transthyretin-mediated amyloidosis. Moreover, the two COVID-19 mRNA vaccines approved by the US FDA are also developed based on LNPs. However, the LNP nucleic acid delivery system still faces several challenges, including the main limitation of the target tissue of LNPs to the liver and the possible immunogenicity.

[0004] LNPs usually consist of four lipid compounds, namely ionizable lipids, neutral lipids, steroids, and PEG-lipids. Among them, the selection of ionizable lipids has the greatest impact on LNPs, such as affecting the encapsulation rate of nucleic acid drugs, the delivery site and efficiency of nucleic acid drugs in vivo, and cytotoxicity.

[0005] Therefore, it is of great significance to develop a novel ionizable lipid with non-liver targeting and biodegradable function. Summary of the Invention

[0006] Problems to be Solved by the Invention

[0007] The first object of this application is to provide a series of compounds, which can be used together with other lipid compounds to prepare lipid carriers, improve the in vivo delivery efficiency of nucleic acid drugs, and specific lipid compounds with a specific structure can be selected as lipid carriers according to the organ where the nucleic acid drug needs to be enriched.

[0008] The second object of this application is to provide a lipid carrier containing the above compounds.

[0009] The third object of the present application is to provide a nucleic acid-lipid nanoparticle composition comprising the above-mentioned compound or the above-mentioned lipid carrier.

[0010] The fourth object of the present application is to provide a pharmaceutical preparation comprising the above-mentioned compound, or the above-mentioned lipid carrier, or the above-mentioned nucleic acid-lipid nanoparticle composition.

[0011] Specific Technical Solution of the Invention

[0012] In one embodiment, the present application provides an ionizable lipid compound represented by the general formula (I), or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, wherein Y, G1, G2, G3, G4, G5, L1, L2, L3, L4, L5, L6, R3 and R4 are as defined in the present application or elsewhere.

[0013]

[0014] In one embodiment, the present application provides a nanoparticle composition comprising the compound provided by the present application and a therapeutic or prophylactic agent. In one embodiment, the therapeutic or prophylactic agent comprises at least one mRNA encoding an antigen or a fragment or epitope thereof.

[0015] Specifically, the present application adopts the following technical solutions.

[0016] 1. A compound of formula (I):

[0017]

[0018] or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, wherein:

[0019] G1 and G2 are each independently an optionally substituted C2-C24 straight-chain alkylene group;

[0020] L1 and L2 are each independently -OC(=O)-, -C(=O)O-, -OC(=O)O-, -C(=O)-, -O-, -S(O) x -, -SS-, -C(=O)S-, -SC(=O)-, -NR a C(=O)-, -C(=O)NR b -, -NR a C(=O)NR b -, -OC(=O)NR b -, -NR a C(=O)O-, -SC(=S)-, -C(=S)S-, -C(=S)-, -CH(OH)-, -P(=O)(OR b) O-, -(C6-C10 arylene)-, or -(6- to 10-membered heteroarylene)-;

[0021] R a and R b are each independently H, optionally substituted C1-C12 alkyl, or optionally substituted C1-C12 alkenyl;

[0022] Said G3 and G4 are each independently optionally substituted C2-C24 straight-chain alkylene;

[0023] Said L3 and L4 are each independently -OC(=O)-, -C(=O)O-, -OC(=O)O-, -C(=O)-, -O-, -S(O) x -, -SS-, -C(=O)S-, -SC(=O)-, -NR c C(=O)-, -C(=O)NR c -, -NR c C(=O)NR d -, -OC(=O)NR c -, -NR c C(=O)O-, -SC(=S)-, -C(=S)S-, -C(=S)-, -CH(OH)-, -P(=O)(OR c )O-, -(C6-C10 arylene)-, or -(6- to 10-membered heteroarylene)-;

[0024] R c and R d are each independently H, optionally substituted C1-C12 alkyl, or optionally substituted C1-C12 alkenyl;

[0025] x is 0, 1, or 2;

[0026] Said G6 and G7 are each independently optionally substituted C2-C24 straight-chain alkylene;

[0027] Said L5 and L6 are each independently -OC(=O)R e , -C(=O)OR e , -OC(=O)OR e , -C(=O)R e , -OR e , -S(O) x R e , -S-SR e , -C(=O)SR e , -SC(=O)R e , -NR e C(=O)R f , -C(=O)NRe R f ,-NR a C(=O)NR e R f ,-OC(=O)NR e R f ,-NR e C(=O)OR f ,-SC(=S)R e ,-C(=S)SR e ,-C(=S)R e ,-CH(OH)R e ,-P(=O)(OR e )(OR f ),-(C6-C10 arylene)-R e ,-(6-to 10-membered heteroarylene)-R e or R e ;

[0028] R e and R f each independently is H, optionally substituted C1-C12 alkyl or optionally substituted C1-C12 alkenyl;

[0029] Said G5 is optionally substituted C1-C12 alkylene;

[0030] R3 is optionally substituted C1-C12 alkyl, optionally substituted C2-C12 alkenyl, optionally substituted C2-C12 alkynyl, optionally substituted C6-C10 aryl or 5-to 10-membered heteroaryl; or R3 and R4 together with the nitrogen to which they are attached form a ring; or R3 is G1L1G3L3L5;

[0031] R4 is optionally substituted C1-C12 alkyl, optionally substituted C2-C12 alkenyl, optionally substituted C2-C12 alkynyl, optionally substituted C6-C10 aryl or 5-to 10-membered heteroaryl; or R4, G5 or a part of G5 together with the nitrogen to which they are attached form a ring; or R4 is G1L1G3L3L5.

[0032] 2. The compound according to item 1, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, wherein the compound has the structure shown in formula (I-A):

[0033]

[0034] wherein, G1, G2, G3, G4, G5, G6, G7, L3, L4, L5, L6, R3 and R4 are as defined in item 1.

[0035] 3. The compound according to item 1, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, wherein the compound has the structure shown in formula (I-B):

[0036]

[0037] wherein the definitions of G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, L4, L5, L6 and R3 are as defined in item 1.

[0038] 4. The compound according to any one of items 1 to 3, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, wherein G1 and G2 are unsubstituted C2-C4 alkylene groups.

[0039] 5. The compound according to any one of items 1 to 4, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, wherein

[0040] G3 and G4 are unsubstituted C2-C4 alkylene groups; or

[0041] wherein G6 and G7 are unsubstituted C2-C4 alkylene groups.

[0042] 6. The compound according to any one of items 1 to 5, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, wherein L1 and L2 are -C(=O)O-.

[0043] 7. The compound according to any one of items 1 to 6, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, wherein L3 and L4 are -OC(=O)-.

[0044] 8. The compound according to any one of items 1 to 7, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, wherein both L5 and L6 are -C(=O)NR e R f .

[0045] 9. The compound according to item 2, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, wherein the compound has the structure shown in formula (I-A-1):

[0046]

[0047] wherein m and n are each independently 1 or 3, and the definitions of R e and R f are as defined in item 1.

[0048] 10. The compound according to item 2, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, characterized in that the compound has the structure shown in formula (I-A-2):

[0049]

[0050] Wherein, m and n are each independently 1 or 3, and R5 and R6 are optionally substituted C1-C12 alkyl, optionally substituted C2-C12 alkenyl, optionally substituted C2-C12 alkynyl, optionally substituted C6-C10 aryl or 5- to 10-membered heteroaryl; or R5 and R6 together with the nitrogen to which they are attached form a ring, and the definitions of G5, R e and R f are as defined in item 1.

[0051] 11. The compound according to item 3, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, characterized in that the compound has the structure shown in formula (I-B-1):

[0052]

[0053] Wherein, m and n are each independently 1 or 3, and the definitions of R e and R f are as defined in item 1.

[0054] 12. The compound according to any one of items 1 to 12, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, wherein R e and / or R f has one of the following structures:

[0055]

[0056] 13. The compound according to any one of items 1 to 12, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, which is a compound in Table 1 below.

[0057] 14. A composition comprising: a therapeutic or prophylactic agent; and a carrier for delivering the therapeutic or prophylactic agent, wherein the carrier comprises an ionizable lipid, and the ionizable lipid comprises one or more of the compounds of formula (I) described in any one of items 1 to 13, or a pharmaceutically available salt thereof.

[0058] 15. The composition according to item 14, wherein the therapeutic or prophylactic agent is selected from one or more of nucleic acid molecules, small molecule compounds, polypeptides or proteins.

[0059] 16. The composition according to item 15, wherein the nucleic acid molecule is selected from one or more of single-stranded DNA, double-stranded DNA, short isoform, agomir, antagomir, antisense molecule, small interfering RNA (siRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), Dicer substrate RNA (dsRNA), short hairpin RNA (shRNA), transfer RNA (tRNA), messenger RNA (mRNA), locked nucleic acid (LNA), peptide nucleic acid (PNA), or morpholino oligonucleotide.

[0060] 17. The composition according to item 16, wherein the therapeutic or prophylactic agent comprises at least one mRNA encoding an antigen or a fragment or epitope thereof.

[0061] 18. The composition according to item 17, wherein the mRNA is monocistronic mRNA or polycistronic mRNA.

[0062] 19. The composition according to item 17, wherein the antigen is a pathogenic antigen.

[0063] 20. The composition according to item 16, wherein the mRNA comprises one or more functional nucleotide analogs selected from one or more of pseudouridine, 1-methyl-pseudouridine, or 5-methylcytosine.

[0064] 21. The composition according to item 15, wherein the small molecule compound is selected from one or more of anti-tumor drugs, anti-infective drugs, local anesthetics, antidepressants, anti-convulsants, antibiotics / antibacterials, anti-fungal drugs, anti-parasitic drugs, hormones, hormone antagonists, immunomodulators, neurotransmitter antagonists, anti-glaucoma agents, anesthetics, or imaging agents.

[0065] 22. The composition according to item 14, wherein the mass ratio of the carrier to the therapeutic or prophylactic agent is 5:1 to 50:1; or;

[0066] The encapsulation rate of the carrier for the therapeutic or prophylactic agent is 50% to 100%.

[0067] 23. The composition according to item 14, wherein the composition is a nanoparticle preparation, and the average size of the nanoparticle preparation is 10 to 500 nm; or the polydispersity index of the nanoparticle preparation is ≤0.5; or the pKa of the nanoparticle preparation is 4.5 to 8.5.

[0068] 24. The composition according to any one of items 14 - 23, wherein the carrier further comprises one or more neutral lipids.

[0069] 25. The composition according to item 24, wherein the neutral lipid is selected from one or more of phosphatidylcholine, phosphatidylethanolamine, sphingomyelin, ceramide, sterol, and its derivatives.

[0070] 26. The composition according to any one of items 14 to 23, wherein the molar ratio of the ionizable lipid to the neutral lipid is 100:1 to 5:1.

[0071] 27. The composition according to any one of items 14 to 23, wherein the carrier further comprises a steroid.

[0072] 28. The composition according to item 27, wherein the steroid is selected from one or more of cholesterol, non-sterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatine, tomatidine, ursolic acid, α-tocopherol, and corticosteroid.

[0073] 29. The composition according to any one of items 14 to 23, wherein the molar ratio of the ionizable lipid to the steroid is 2:1 to 4:1.

[0074] 30. The composition according to any one of items 14 to 23, wherein the carrier further comprises one or more lipids capable of binding to a polymer.

[0075] 31. The composition according to item 30, wherein the lipid capable of binding to a polymer is selected from one or more of PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, and PEG-modified dialkylglycerol.

[0076] 32. The composition according to item 30 or 31, wherein the molar ratio of the ionizable lipid to the lipid capable of binding to a polymer is 100:1 to 20:1.

[0077] 33. The composition according to any one of items 14 to 23, the carrier further comprises a neutral lipid, a structural lipid, and a polymer-conjugated lipid, and the molar ratio of the ionizable lipid, the neutral lipid, the steroid lipid, and the polymer-conjugated lipid is (15 to 70):(1 to 15):(15 to 35):(0 to 3).

[0078] 34. The composition according to item 14, characterized in that the composition further comprises a pharmaceutically acceptable excipient.

[0079] 35. The composition according to item 34, characterized in that the excipient comprises a pharmaceutically acceptable diluent.

[0080] 36. Use of a compound represented by formula (I) as described in any one of items 1 to 13, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, or the composition described in any one of items 14 to 35 in the preparation of a drug, preferably the drug is any one selected from gene drugs, nucleic acid vaccines, small molecule drugs, polypeptides or protein drugs.

[0081] Those skilled in the art will recognize additional features of the disclosure after considering the following detailed description of specific embodiments.

[0082] Advantages of the Invention

[0083] The ionizable lipid compound of the present application can be used to deliver nucleic acid drugs or small molecule drugs, enriching the types of ionizable lipid compounds, which is of great significance for the development and application of nucleic acid prophylactic and therapeutic agents. Brief Description of the Drawings

[0084] Figure 1 Relative fluorescence intensity of mFluc expressed in the spleen delivered by each lipid nanoparticle;

[0085] Figure 2 Relative fluorescence intensity of fluorescent protein mRNA delivered to each organ by A1-H226x lipid nanoparticles;

[0086] Figure 3 Effect of changes in the molar ratio of DSPC and cholesterol on the delivery efficiency;

[0087] Figure 4 Effect of changes in the molar ratio of ionizable lipid and DMG-PEG2000 on the delivery efficiency;

[0088] Figure 5 Effect of different phospholipids on the delivery efficiency;

[0089] Figure 6 Analysis of IgG antibody levels in the spleen of tumor model mice;

[0090] Figure 7 Results of immune cell typing in the tumor tissue of tumor model mice;

[0091] Figure 8 Analysis of antigen-specific T cells in the tumor tissue of tumor model mice;

[0092] Figure 9 Analysis of T cells in the tumor tissue of tumor model mice;

[0093] Figure 10 Analysis of T cells in the spleen cells of tumor model mice;

[0094] Figure 11 Inhibitory effect of lipid nanoparticle vaccine encapsulating antigen on tumor growth. Detailed Implementation Modes

[0095] Terms

[0096] Unless otherwise described, all technical and scientific terms used in this application have the same meaning as commonly understood by those skilled in the art. For the purpose of interpreting this specification, the following terms will be applied, and where appropriate, terms used in the singular form will also include the plural form, and vice versa. All patents, applications, published applications, and other publications are incorporated by reference in their entirety. If any description of the terms conflicts with any document incorporated by reference into this application, the description of the terms set forth below shall prevail.

[0097] Unless the context otherwise requires, in this specification and the claims, the word "comprise" and its variations, such as "comprising" and "containing", are interpreted in an open and inclusive sense, i.e., "including, but not limited to".

[0098] As used in this application and unless otherwise specified, the term "lipid" refers to a group of organic compounds that includes, but is not limited to, fatty acid esters, and is generally characterized by being poorly soluble in water but soluble in many nonpolar organic solvents. Although lipids generally have weak water solubility, certain classes of lipids (such as lipids modified with polar groups, such as DMG-PEG2000) have limited water solubility and are soluble in water under certain conditions. Known lipid types include biomolecules such as fatty acids, waxes, sterols, fat-soluble vitamins, monoglycerides, diglycerides, triglycerides, and phospholipids. Lipids can be classified into at least three categories: (1) "simple lipids", including fats and oils, and waxes; (2) "compound lipids", including phospholipids and glycolipids (such as DMPEPEG2000); and (3) "derived lipids", such as steroids. In addition, as used in this application, lipids also include lipid-like compounds. The term "lipid-like compound" is also abbreviated as "lipoid", and refers to lipid-like compounds (such as amphiphilic compounds with lipid-like physical properties).

[0099] As used in this application and unless otherwise specified, the term "lipid nanoparticle" or "LNP" refers to particles having at least one dimension in the nanometer (nm) range (e.g., 1 to 1,000 nm) that contain one or more types of lipid molecules. The LNPs provided in this application may further contain at least one non-lipid payload molecule (e.g., one or more nucleic acid molecules) or small molecule drugs. In some embodiments, the LNP contains a non-lipid payload molecule that is partially or fully encapsulated within a lipid shell. Where the payload is a negatively charged molecule (e.g., mRNA encoding a viral protein), and the lipid component of the LNP contains at least one ionizable lipid. Without being bound by theory, it is expected that the ionizable lipid can interact with the negatively charged payload molecule and facilitate incorporation and / or encapsulation of the payload into the LNP during LNP formation. Other lipids that can form part of the LNPs provided in this application include, but are not limited to, neutral lipids and charged lipids such as steroids or their analogs, polymer-conjugated lipids, and various zwitterionic lipids.

[0100] As used in this application and unless otherwise specified, the term "ionizable lipid" refers to a lipid that is capable of being ionized to a positively charged form. Ionizable lipids include one or more amine groups that can form a positive charge so that they can exist in a positively charged form depending on the pH value. The ionization of the ionizable lipid affects the surface charge of the lipid nanoparticle under different conditions. This charge state can affect plasma protein uptake, blood clearance, and tissue distribution (Semple, S.C. et al., Adv Drug Deliv Rev 32:3-17 (1998)) as well as the ability to form endosomolytic non-bilayer structures (Hafez, I.M. et al., Gene Ther 8:1188-1196 (2001)), which is crucial for intracellular delivery of nucleic acids.

[0101] As used in this application and unless otherwise specified, the term "steroid" refers to a compound containing the following carbon skeleton:

[0102]

[0103] Non-limiting examples of steroids include cholesterol and the like.

[0104] As used in this application and unless otherwise specified, the term "neutral lipid" encompasses any lipid molecule that exists in an uncharged form or in a neutral zwitterionic form at a selected pH value or within a selected pH range. In some embodiments, the selected useful pH value or range corresponds to the pH conditions in the environment of a predetermined lipid use, such as physiological pH. As non-limiting examples, neutral lipids that can be used in conjunction with the present disclosure include, but are not limited to, phosphatidylcholines, such as 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC); phosphatidylethanolamines, such as 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 2-((2,3-bis(oleoyloxy)propyl))dimethylammonio)ethyl hydrogen phosphate (DOCP); sphingomyelin (SM); ceramides; steroids, such as sterols, phytosterols, and their derivatives. The neutral lipids provided in this application can be synthetic or derived from natural sources or compounds (from which they are isolated or modified).

[0105] As used in this application and unless otherwise specified, the term "alkyl" refers to a saturated straight-chain or branched hydrocarbon chain group consisting only of carbon and hydrogen atoms. In one embodiment, the alkyl has, for example, one to twenty-four carbon atoms (C1-C24 alkyl), four to twenty carbon atoms (C4-C20 alkyl), six to sixteen carbon atoms (C6-C16 alkyl), six to nine carbon atoms (C6-C9 alkyl), one to fifteen carbon atoms (C1-C15 alkyl), one to twelve carbon atoms (C1-C12 alkyl), one to eight carbon atoms (C1-C8 alkyl), or one to six carbon atoms (C1-C6 alkyl) and is attached to the remainder of the molecule by a single bond. Examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, 1,1-dimethylethyl (tert-butyl), 3-methylhexyl, 2-methylhexyl, and the like. Unless otherwise specified, the alkyl is optionally substituted.

[0106] As used in this application and unless otherwise indicated, the term "alkenyl" refers to a straight or branched hydrocarbon chain group consisting only of carbon and hydrogen atoms, which contains one or more carbon-carbon double bonds. Those skilled in the art will understand that the term "alkenyl" also includes groups having "cis" and "trans" configurations, or having "E" and "Z" configurations. In one embodiment, the alkenyl has, for example, two to twenty-four carbon atoms (C2-C24 alkenyl), four to twenty carbon atoms (C4-C20 alkenyl), six to sixteen carbon atoms (C6-C16 alkenyl), six to nine carbon atoms (C6-C9 alkenyl), two to fifteen carbon atoms (C2-C15 alkenyl), two to twelve carbon atoms (C2-C12 alkenyl), two to eight carbon atoms (C2-C8 alkenyl) or two to six carbon atoms (C2-C6 alkenyl) and is connected to the rest of the molecule by a single bond. Examples of alkenyl include, but are not limited to, vinyl, prop-1-enyl, but-1-enyl, pent-1-enyl, pent-1,4-dienyl, etc. Unless otherwise indicated, the alkenyl is optionally substituted.

[0107] As used in this application and unless otherwise indicated, the term "alkynyl" refers to a straight or branched hydrocarbon chain group consisting only of carbon and hydrogen atoms, which contains one or more carbon-carbon triple bonds. In one embodiment, the alkynyl has, for example, two to twenty-four carbon atoms (C2-C24 alkynyl), four to twenty carbon atoms (C4-C20 alkynyl), six to sixteen carbon atoms (C6-C16 alkynyl), six to nine carbon atoms (C6-C9 alkynyl), two to fifteen carbon atoms (C2-C15 alkynyl), two to twelve carbon atoms (C2-C12 alkynyl), two to eight carbon atoms (C2-C8 alkynyl) or two to six carbon atoms (C2-C6 alkynyl) and is connected to the rest of the molecule by a single bond. Examples of alkynyl include, but are not limited to, ethynyl, propynyl, butynyl, pentynyl, etc. Unless otherwise indicated, the alkynyl is optionally substituted.

[0108] As used in this application and unless otherwise indicated, the term "cyclizing" refers to the local or complete connection within or between molecules to form a cyclic molecular structure, and the connection points can be made by including, but not limited to, one C or N or several C or N. The cyclized molecular structure can be saturated or unsaturated. Unless otherwise indicated, the cyclized molecular moiety is optionally substituted.

[0109] When a group described in the present application is referred to as "substituted", it may be substituted with one or more any suitable substituents. Illustrative examples of substituents include, but are not limited to, substituents found in the exemplary compounds and embodiments provided in the present application, and: halogen atoms such as F, Cl, Br or I; cyano; oxo (=O); hydroxy (-OH); alkyl; alkenyl; alkynyl; cycloalkyl; aryl; -(C=O)OR'; -O(C=O)R'; -C(=O)R'; -OR'; -S(O) x R'; -S-SR'; -C(=O)SR'; -SC(=O)R'; -NR'R'; -NR'C(=O)R'; -C(=O)NR'R'; -NR'C(=O)NR'R'; -OC(=O)NR'R'; -NR'C(=O)OR'; -NR'S(O) x NR'R'; -NR'S(O) x R'; and -S(O) x NR'R', where: R' is independently H, C1-C15 alkyl or cycloalkyl each time it appears, and x is 0, 1 or 2.

[0110] As used in the present application and unless otherwise specified, the term "optionally selected" or "optionally" (e.g., optionally substituted) means that the subsequently described event or circumstance may or may not occur, and the description includes both the case where the event or circumstance occurs and the case where it does not occur. For example, "optionally substituted alkyl" means that the alkyl may be substituted or may not be substituted, and the description includes both substituted alkyl and unsubstituted alkyl.

[0111] The term "prodrug" refers to a derivative that can directly or indirectly provide the compounds or nucleic acid drugs of the present application after being administered to a patient. Prodrugs of a composition can be prepared by modifying the functional groups present in the compound or nucleic acid (DNA, ASO, siRNA, mRNA, tRNA, saRNA) in such a way that the modification can be cleaved in a conventional operation or in vivo to obtain the parent compound or nucleic acid. Particularly preferred prodrugs are compounds and nucleic acid drugs that can improve the bioavailability of the compositions of the present application (e.g., are more readily absorbed into the blood) when administered to a patient, or compounds and nucleic acid drugs that facilitate the delivery of the parent compound to the site of action (e.g., the lymphatic system). Unless otherwise indicated, all prodrug forms of the compounds of the present application are within the scope of the present application, and various prodrug forms are well known in the art.

[0112] As used in the present application and unless otherwise specified, the term "pharmaceutically acceptable salt" includes both acid addition salts and base addition salts.

[0113] Examples of pharmaceutically acceptable acid addition salts include, but are not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.; and organic acids such as, but not limited to, acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, camphoric acid, camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecyl sulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronic acid, glutamic acid, glutaric acid, 2-oxoglutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, pyroglutamic acid, pyruvic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid, undecylenic acid, etc.

[0114] Examples of pharmaceutically acceptable base addition salts include, but are not limited to, salts prepared by adding an inorganic base or an organic base to a free acid compound. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts, etc. In one embodiment, the inorganic salts are ammonium salts, sodium salts, potassium salts, calcium salts, and magnesium salts. Salts derived from organic bases include, but are not limited to, salts of primary amines, secondary amines, and tertiary amines; substituted amines, including naturally occurring substituted amines; cyclic amines, and basic ion exchange resins such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, deanol, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, benethamine, benzathine, ethylenediamine, glucosamine, methylglucosamine, theobromine, triethanolamine, tromethamine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins, etc.

[0115] The compounds provided by the present application may contain one or more asymmetric centers and may thus give rise to enantiomers, diastereomers, and other stereoisomeric forms, which may be defined as (R)- or (S)- according to absolute stereochemistry or as (D)- or (L)- for amino acids. Unless otherwise specified, the compounds provided by the present application are intended to include all such possible isomers, as well as their racemic and optically pure forms. When the compounds described in the present application contain an olefinic double bond or other geometrically asymmetric center, unless otherwise specified, the compounds are intended to include E and Z geometric isomers. Similarly, all tautomeric forms are also intended to be included.

[0116] As used in the present application and unless otherwise specified, the term "isomer" refers to different compounds having the same molecular formula. "Stereoisomers" are isomers that differ only in the arrangement of atoms in space. "Atropisomers" are stereoisomers obtained by restricted rotation about a single bond. "Enantiomers" are a pair of stereoisomers that are non-superimposable mirror images of each other. A mixture of any ratio of a pair of enantiomers may be referred to as a "racemic" mixture. "Diastereomers" are stereoisomers that have at least two asymmetric atoms but are not mirror images of each other.

[0117] "Stereoisomers" may also include E and Z isomers or mixtures thereof, as well as cis and trans isomers or mixtures thereof. In certain embodiments, the compounds described in the present application are separated into E or Z isomers. In other embodiments, the compounds described in the present application are a mixture of E and Z isomers.

[0118] The term "nucleic acid" refers to a polymer of nucleotides of any length and includes, for example, DNA and RNA. The nucleotides can be deoxyribonucleotides, ribonucleotides, modified nucleotides or bases and / or their analogs, or any substrate that can be incorporated into the polymer by DNA or RNA polymerase or by a synthetic reaction. The nucleic acid can be in single-stranded or double-stranded form. As used in this application and unless otherwise specified, "nucleic acid" also includes nucleic acid mimetics such as locked nucleic acid (LNA), peptide nucleic acid (PNA), and morpholino nucleic acid. As used in this application, "oligonucleotide" refers to a short synthetic polynucleotide, generally but not necessarily less than about 200 nucleotides in length. The terms "oligonucleotide" and "polynucleotide" are not mutually exclusive. The above description of polynucleotides applies equally and fully to oligonucleotides. Unless otherwise specified, the left-hand end of any single-stranded polynucleotide sequence disclosed in this application is the 5' end; the left-hand direction of a double-stranded polynucleotide sequence is called the 5' direction. The 5' to 3' addition direction of nascent RNA transcripts is called the transcription direction; the sequence region on the DNA strand that has the same sequence as the RNA transcript and is located at the 5' end relative to the 5' end of the RNA transcript is called the "upstream sequence"; the sequence region on the DNA strand that has the same sequence as the RNA transcript and is located at the 3' end relative to the 3' end of the RNA transcript is called the "downstream sequence".

[0119] "Isolated nucleic acid" refers to a nucleic acid that is substantially separated from other genomic DNA sequences and proteins or complexes (such as ribosomes and polymerases) that naturally accompany the native sequence, such as RNA, DNA, or hybrid nucleic acids. An "isolated" nucleic acid molecule is a nucleic acid molecule that is separated from other nucleic acid molecules present in the natural source of the nucleic acid molecule. In addition, when produced by recombinant techniques, an "isolated" nucleic acid molecule, such as an mRNA molecule, can be substantially free of other cellular material or medium, or when chemically synthesized, it can be substantially free of chemical precursors or other chemicals. In certain embodiments, one or more nucleic acid molecules encoding an antigen described in this application are isolated or purified. The term includes nucleic acid sequences that have been removed from their natural environment and includes recombinant or cloned DNA or RNA isolates as well as chemically synthesized analogs or analogs biosynthesized by heterologous systems. Substantially pure molecules can include isolated forms of the molecule.

[0120] The term "coding nucleic acid" or its grammatical equivalent, when used to refer to a nucleic acid molecule, includes: (a) a nucleic acid molecule that, when in its native state or when manipulated by methods well known to those of skill in the art, can be transcribed to produce mRNA and then translated into a peptide and / or polypeptide; and (b) the mRNA molecule itself. The antisense strand is the complementary sequence of such a nucleic acid molecule, and the coding sequence can be deduced therefrom. The term "coding region" refers to the portion of a coding nucleic acid sequence that is translated into a peptide or polypeptide. The term "untranslated region" or "UTR" refers to the portion of a coding nucleic acid that is not translated into a peptide or polypeptide. Depending on the orientation of the UTR relative to the coding region of the nucleic acid molecule, the UTR is called a 5'-UTR if it is located at the 5' end of the coding region, and a 3'-UTR if it is located at the 3' end of the coding region.

[0121] As used in this application, the term "mRNA" refers to a messenger RNA molecule that contains one or more open reading frames (ORFs) that can be translated by a cell or organism having the mRNA to produce one or more peptide or protein products. The region containing one or more ORFs is called the coding region of the mRNA molecule. In certain embodiments, the mRNA molecule further contains one or more untranslated regions (UTRs).

[0122] In certain embodiments, the mRNA is a monocistronic mRNA that contains only one ORF. In certain embodiments, the monocistronic mRNA encodes a peptide or protein that contains at least one epitope of a selected antigen (e.g., a pathogenic antigen or a tumor-associated antigen). In other embodiments, the mRNA is a polycistronic mRNA that contains two or more ORFs. In certain embodiments, the polycistronic mRNA encodes two or more peptides or proteins that can be the same or different from each other. In certain embodiments, each peptide or protein encoded by the polycistronic mRNA contains at least one epitope of a selected antigen. In certain embodiments, the different peptides or proteins encoded by the polycistronic mRNA each contain at least one epitope of a different antigen. In any of the embodiments described in this application, the at least one epitope can be at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 epitopes of an antigen.

[0123] The term "nucleobase" encompasses purines and pyrimidines, including the natural compounds adenine, thymine, guanine, cytosine, uracil, inosine, and their natural or synthetic analogs or derivatives.

[0124] As used herein, the term "functional nucleotide analogue" refers to a modified form of a classical nucleotide A, G, C, U or T, said form (a) retaining the base pairing properties of the corresponding classical nucleotide and (b) containing at least one chemical modification to the corresponding native nucleotide of (i) the nucleobase, (ii) the glycosyl group, (iii) the phosphate group or (iv) any combination of (i) to (iii). As used herein, base pairing encompasses not only the classical Watson-Crick adenine-thymine, adenine-uracil or guanine-cytosine base pairs, but also base pairs formed between a classical nucleotide and a functional nucleotide analogue or between a pair of functional nucleotide analogues, where the arrangement of hydrogen bond donors and hydrogen bond acceptors permits the formation of hydrogen bonds between the modified nucleobase and the classical nucleobase or between two complementary modified nucleobase structures. For example, a functional analogue of guanosine (G) retains the ability to base pair with cytosine (C) or a functional analogue of cytosine. An example of such non-classical base pairing is the base pairing between the modified nucleotide inosine and adenine, cytosine or uracil. As used herein, functional nucleotide analogues can be naturally occurring or non-naturally occurring. Thus, a nucleic acid molecule containing a functional nucleotide analogue can have at least one modified nucleobase, glycosyl group and / or internucleoside linkage. Exemplary chemical modifications to the nucleobase, glycosyl group or internucleoside linkage of a nucleic acid molecule are provided herein.

[0125] As used herein, the terms "translation enhancer element", "TEE" and "translation enhancer" refer to a region in a nucleic acid molecule that functions to promote the translation of the coding sequence of the nucleic acid into a protein or peptide product, such as by cap-dependent or cap-independent translation. TEEs are typically located in the UTR region of a nucleic acid molecule (e.g., mRNA) and enhance the level of translation of a coding sequence located either upstream or downstream. For example, a TEE in the 5'-UTR of a nucleic acid molecule can be located between the promoter and the start codon of the nucleic acid molecule. A variety of TEE sequences are known in the art (Wellensiek et al., Genome-wide profiling of human cap-independent translation-enhancing elements, Nature Methods, August 2013; 10(8):747-750; Chappell et al., PNAS, June 29, 2004, 101(26)9590-9594). Some TEEs are known to be conserved across multiple species (Pánek et al., Nucleic Acids Research, Vol. 41, No. 16, September 1, 2013, pp. 7625-7634).

[0126] As used in this application, the term "peptide" refers to a polymer containing two to fifty (2 - 50) amino acid residues joined by one or more covalent peptide bonds. The term applies to both naturally occurring amino acid polymers and amino acid polymers in which one or more of the amino acid residues are non - naturally occurring amino acids (such as amino acid analogs or unnatural amino acids).

[0127] The terms "polypeptide" and "protein" are used interchangeably in this application and refer to a polymer having more than fifty (50) amino acid residues joined by covalent peptide bonds. That is, descriptions of polypeptides apply equally to descriptions of proteins and vice versa. The term applies to both naturally occurring amino acid polymers and amino acid polymers in which one or more of the amino acid residues are non - naturally occurring amino acids (such as amino acid analogs). As used in this application, the term encompasses amino acid chains of any length, including full - length proteins (such as antigens).

[0128] The term "antigen" refers to a substance that can be recognized by the immune system of a subject (including the adaptive immune system) and can trigger an immune response (including an antigen - specific immune response) after contact between the subject and the antigen. In certain embodiments, the antigen is a protein associated with diseased cells, such as cells infected with a pathogen or neoplastic cells (e.g., tumor - associated antigen (TAA)).

[0129] An "epitope" is the site on the surface of an antigen molecule that binds to a single antibody molecule, such as a local region on the antigen surface that can bind to one or more antigen - binding regions of an antibody, and that has antigenic or immunogenic activity in an animal, such as in a mammal (e.g., in a human), capable of eliciting an immune response. An epitope with immunogenic activity is the part of a polypeptide that elicits an antibody response in an animal. An epitope with antigenic activity is the part of a polypeptide that is bound by an antibody determined by any method well - known in the art, including, for example, by immunoassay. An antigenic epitope does not necessarily have immunogenicity. Epitopes are usually composed of chemically active surface groups of a molecule, such as amino acids or sugar side - chains, and have specific three - dimensional structural features as well as specific charge features. Antibody epitopes can be linear epitopes or conformational epitopes. A linear epitope is formed by a continuous amino acid sequence in a protein. A conformational epitope is formed by amino acids that are not contiguous in the protein sequence but that come together when the protein folds into its three - dimensional structure. An induced epitope is formed when the three - dimensional structure of a protein assumes an altered conformation, such as after activation or binding by another protein or ligand. In certain embodiments, an epitope is a three - dimensional surface feature of a polypeptide. In other embodiments, an epitope is a linear feature of a polypeptide. Generally, an antigen has several or many different epitopes and can react with many different antibodies.

[0130] Lipid composition

[0131] In one embodiment, the present application provides a compound of formula (I):

[0132]

[0133] or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, wherein:

[0134] each of G1 and G2 is independently an optionally substituted C2-C24 straight-chain alkylene group;

[0135] each of L1 and L2 is independently -OC(=O)-, -C(=O)O-, -OC(=O)O-, -C(=O)-, -O-, -S(O) x -, -SS-, -C(=O)S-, -SC(=O)-, -NR a C(=O)-, -C(=O)NR b -, -NR a C(=O)NR b -, -OC(=O)NR b -, -NR a C(=O)O-, -SC(=S)-, -C(=S)S-, -C(=S)-, -CH(OH)-, -P(=O)(OR b )O-, -(C6-C10 arylene)- or -(6- to 10-membered heteroarylene)-;

[0136] R a and R b are each independently H, an optionally substituted C1-C12 alkyl group or an optionally substituted C1-C12 alkenyl group;

[0137] each of G3 and G4 is independently an optionally substituted C2-C24 straight-chain alkylene group;

[0138] each of L3 and L4 is independently -OC(=O)-, -C(=O)O-, -OC(=O)O-, -C(=O)-, -O-, -S(O) x -, -SS-, -C(=O)S-, -SC(=O)-, -NR c C(=O)-, -C(=O)NR c -, -NR c C(=O)NR d -, -OC(=O)NR c -, -NR c C(=O)O-, -SC(=S)-, -C(=S)S-, -C(=S)-, -CH(OH)-, -P(=O)(OR c) O-, -(C6-C10 arylene)-, or -(6 to 10-membered heteroarylene)-;

[0139] R c and R d are each independently H, optionally substituted C1-C12 alkyl, or optionally substituted C1-C12 alkenyl;

[0140] x is 0, 1, or 2;

[0141] said G6 and G7 are each independently optionally substituted C2-C24 straight-chain alkylene;

[0142] said L5 and L6 are each independently -OC(=O)R e , -C(=O)OR e , -OC(=O)OR e , -C(=O)R e , -OR e , -S(O) x R e , -S-SR e , -C(=O)SR e , -SC(=O)R e , -NR e C(=O)R f , -C(=O)NR e R f , -NR a C(=O)NR e R f , -OC(=O)NR e R f , -NR e C(=O)OR f , -SC(=S)R e , -C(=S)SR e , -C(=S)R e , -CH(OH)R e , -P(=O)(OR e )(OR f ), -(C6-C10 arylene)-R e , -(6 to 10-membered heteroarylene)-R e or R e ;

[0143] R e and R f are each independently H, optionally substituted C1-C12 alkyl, or optionally substituted C1-C12 alkenyl;

[0144] said G5 is optionally substituted C1-C12 alkylene;

[0145] R3 is an optionally substituted C1-C12 alkyl group, an optionally substituted C2-C12 alkenyl group, an optionally substituted C2-C12 alkynyl group, an optionally substituted C6-C10 aryl group or a 5- to 10-membered heteroaryl group; or R3 and R4 together with the nitrogen to which they are attached form a ring; or R3 is G1L1G3L3L5;

[0146] R4 is an optionally substituted C1-C12 alkyl group, an optionally substituted C2-C12 alkenyl group, an optionally substituted C2-C12 alkynyl group, an optionally substituted C6-C10 aryl group or a 5- to 10-membered heteroaryl group; or R4, G5 or a part of G5 together with the nitrogen to which they are attached form a ring; or R4 is G1L1G3L3L5.

[0147] Preferably, L1 and L2 are -C(=O)O-, and the compound has the structure of formula (I-A):

[0148]

[0149] wherein the definitions of G1, G2, G3, G4, G5, G6, G7, L3, L4, L5, L6, R3 and R4 are as defined in formula (I).

[0150] Alternatively preferably, R4 is -G1L1G3L3L5, and the compound has the structure shown in formula (I-B):

[0151]

[0152] wherein the definitions of G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, L4, L5, L6, R3 are as defined in formula (I).

[0153] Preferably, G1 and G2 are unsubstituted C2-C4 alkylene groups, wherein the definitions of G3, G4, G5, G6, G7, L1, L2, L3, L4, L5, L6, R3 and R4 are as defined in formula (I).

[0154] Preferably, G3 and G4 are unsubstituted C2-C4 alkylene groups, wherein the definitions of G1, G2, G5, G6, G7, L1, L2, L3, L4, L5, L6, R3 and R4 are as defined in formula (I).

[0155] Preferably, G5 is an unsubstituted C2-C5 alkylene group, wherein the definitions of G1, G2, G3, G4, G6, G7, L1, L2, L3, L4, L5, L6, R3 and R4 are as defined in formula (I).

[0156] Preferably, G6 and G7 are unsubstituted C2-C4 alkylene groups, wherein G1, G2, G3, G4, G5, L1, L2, L3, L4, L5, L6, R3 and R4 are as defined in formula (I).

[0157] Preferably, L1 and L2 are -C(=O)O-, wherein G1, G2, G3, G4, G5, G6, G7, L3, L4, L5, L6, R3 and R4 are as defined in formula (I).

[0158] Preferably, L3 and L4 are -OC(=O)-, wherein G1, G2, G3, G4, G5, G6, G7, L1, L2, L5, L6, R3 and R4 are as defined in formula (I).

[0159] Preferably, both L5 and L6 are -C(=O)NR e R f , wherein G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, L4, R3 and R4 are as defined in formula (I), and R e and R f are both C6-C8 straight-chain alkyl groups.

[0160] Preferably, both L5 and L6 are -C(=O)NR e R f , wherein G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, L4, R3 and R4 are as defined in formula (I), and R e and R f are both C6-C8 branched-chain alkyl groups.

[0161] Preferably, R3 is a C1-C3 alkyl group, wherein G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, L4, L5, L6 and R4 are as defined in formula (I).

[0162] Preferably, R3 is G1L1G3L3L5, wherein G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, L4, L5, L6 and R4 are as defined in formula (I).

[0163] Preferably, R4 is a C1-C3 alkyl group, wherein G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, L4, L5, L6 and R3 are as defined in formula (I).

[0164] Preferably, R4 is a C6-C10 aryl group, wherein the definitions of G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, L4, L5, L6 and R3 are as defined in formula (I).

[0165] Preferably, R4 is a C1-C12 alkyl group, and R4, G5 or a part of G5 together with the nitrogen atom to which it is attached form a ring, wherein the definitions of G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, L4, L5, L6 and R3 are as defined in formula (I).

[0166] Preferably, R3 and R4 are C1-C12 alkyl groups, and R3, R4 together with the nitrogen atom to which they are attached form a ring, wherein the definitions of G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, L4, L5, L6 are as defined in formula (I).

[0167] Preferably, R4 is a C1-C3 alkyl group; R3 is a C1-C3 alkyl group, wherein the definitions of G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, L4, L5, L6 are as defined in formula (I).

[0168] Preferably, R4 is a C6-C10 aryl group; R3 is a C1-C3 alkyl group, wherein the definitions of G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, L4, L5, L6 are as defined in formula (I).

[0169] Preferably, R4 is a C1-C12 alkyl group, and R4, G5 or a part of G5 together with the nitrogen atom to which it is attached form a ring; R3 is a C1-C3 alkyl group, wherein the definitions of G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, L4, L5, L6 are as defined in formula (I).

[0170] Preferably, R4 is a C1-C3 alkyl group; R3 is G1L1G3L3L5, wherein the definitions of G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, L4, L5, L6 are as defined in formula (I).

[0171] Preferably, R4 is a C6-C10 aryl group; R3 is G1L1G3L3L5, wherein the definitions of G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, L4, L5, L6 are as defined in formula (I).

[0172] Preferably, R4 is a C1-C12 alkyl group, and R4, G5 or a portion of G5 together with the nitrogen to which they are connected form a ring; R3 is G1L1G3L3L5, wherein G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, L4, L5, and L6 are defined as in formula (I).

[0173] Preferably, R3 and R4 are C1-C12 alkyl groups, and R3, R4 and the nitrogen to which they are connected form a ring; L5 and L6 are both -C(=O)NR e R f , R e and R f are all C6-C8 branched alkyl groups, wherein G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, and L4 are as defined in formula (I).

[0174] Preferably, R4 is C1-C3 alkyl; R3 is C1-C3 alkyl; L5 and L6 are both -C(=O)NR e R f , R e and R f are all C6-C8 branched alkyl groups, wherein G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, and L4 are as defined in formula (I).

[0175] Preferably, R4 is a C6-C10 aryl group; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f , R e and R f are all C6-C8 branched alkyl groups, wherein G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, and L4 are as defined in formula (I).

[0176] Preferably, R4 is a C1-C12 alkyl group, R4, G5 or a portion of G5 together with the nitrogen to which they are attached form a ring; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f , R e and R f are all C6-C8 branched alkyl groups, wherein G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, and L4 are as defined in formula (I).

[0177] Preferably, R4 is C1-C3 alkyl; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f , R e and Rf All are C6-C8 branched alkyl groups, wherein the definitions of G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, and L4 are as defined in formula (I).

[0178] Preferably, R4 is a C6-C10 aryl group; R3 is G1L1G3L3L5; both L5 and L6 are -C(=O)NR e R f R e and R f All are C6-C8 branched alkyl groups, wherein the definitions of G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, and L4 are as defined in formula (I).

[0179] Preferably, R4 is a C1-C12 alkyl group, R4, G5 or a part of G5 together with the nitrogen to which it is attached form a ring; R3 is G1L1G3L3L5; both L5 and L6 are -C(=O)NR e R f R e and R f All are C6-C8 branched alkyl groups, wherein the definitions of G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, and L4 are as defined in formula (I).

[0180] Preferably, R3 and R4 are C1-C12 alkyl groups, R3 and R4 together with the nitrogen to which they are attached form a ring; both L5 and L6 are -C(=O)NR e R f R e and R f All are C6-C8 straight-chain alkyl groups, wherein the definitions of G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, and L4 are as defined in formula (I).

[0181] Preferably, R4 is a C1-C3 alkyl group; R3 is a C1-C3 alkyl group; both L5 and L6 are -C(=O)NR e R f R e and R f All are C6-C8 straight-chain alkyl groups, wherein the definitions of G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, and L4 are as defined in formula (I).

[0182] Preferably, R4 is a C6-C10 aryl group; R3 is a C1-C3 alkyl group; both L5 and L6 are -C(=O)NR e R f R e and R fAll are straight-chain C6-C8 alkyl groups, where the definitions of G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, and L4 are as defined in formula (I).

[0183] Preferably, R4 is a C1-C12 alkyl group, and R4, G5 or a part of G5 together with the nitrogen to which it is attached form a ring; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f ,R e and R f All are straight-chain C6-C8 alkyl groups, where the definitions of G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, and L4 are as defined in formula (I).

[0184] Preferably, R4 is a C1-C3 alkyl group; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f ,R e and R f All are straight-chain C6-C8 alkyl groups, where the definitions of G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, and L4 are as defined in formula (I).

[0185] Preferably, R4 is a C6-C10 aryl group; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f ,R e and R f All are straight-chain C6-C8 alkyl groups, where the definitions of G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, and L4 are as defined in formula (I).

[0186] Preferably, R4 is a C1-C12 alkyl group, and R4, G5 or a part of G5 together with the nitrogen to which it is attached form a ring; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f ,R e and R f All are straight-chain C6-C8 alkyl groups, where the definitions of G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, and L4 are as defined in formula (I).

[0187] Preferably, R3 and R4 are C1-C12 alkyl groups, and R3, R4 together with the nitrogen to which they are attached form a ring; L5 and L6 are both -C(=O)NR e R f ,R e and R fAll are C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-, where the definitions of G1, G2, G3, G4, G5, G6, G7, L1, and L2 are as defined in formula (I).

[0188] Preferably, R4 is a C1-C3 alkyl group; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f ,R e and R f All are C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-, where the definitions of G1, G2, G3, G4, G5, G6, G7, L1, and L2 are as defined in formula (I).

[0189] Preferably, R4 is a C6-C10 aryl group; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f ,R e and R f All are C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-, where the definitions of G1, G2, G3, G4, G5, G6, G7, L1, and L2 are as defined in formula (I).

[0190] Preferably, R4 is a C1-C12 alkyl group, and R4, G5 or a part of G5 together with the nitrogen to which it is attached form a ring; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f ,R e and R f All are C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-, where the definitions of G1, G2, G3, G4, G5, G6, G7, L1, and L2 are as defined in formula (I).

[0191] Preferably, R4 is a C1-C3 alkyl group; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f ,R e and R f All are C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-, where the definitions of G1, G2, G3, G4, G5, G6, G7, L1, and L2 are as defined in formula (I).

[0192] Preferably, R4 is a C6-C10 aryl group; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f ,R e and R fare all C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-, wherein G1, G2, G3, G4, G5, G6, G7, L1, and L2 are as defined in formula (I).

[0193] Preferably, R4 is a C1-C12 alkyl group, R4, G5 or a portion of G5 together with the nitrogen to which it is attached forms a ring; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f , R e and R f are all C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-, wherein G1, G2, G3, G4, G5, G6, G7, L1, and L2 are as defined in formula (I).

[0194] Preferably, R3 and R4 are C1-C12 alkyl groups, and R3, R4 and the nitrogen to which they are connected form a ring; L5 and L6 are both -C(=O)NR e R f , R e and R f are all C6-C8 straight chain alkyl groups; L3 and L4 are -OC(=O)-, wherein G1, G2, G3, G4, G5, G6, G7, L1 and L2 are as defined in formula (I).

[0195] Preferably, R4 is C1-C3 alkyl; R3 is C1-C3 alkyl; L5 and L6 are both -C(=O)NR e R f , R e and R f are all C6-C8 straight chain alkyl groups; L3 and L4 are -OC(=O)-, wherein G1, G2, G3, G4, G5, G6, G7, L1 and L2 are as defined in formula (I).

[0196] Preferably, R4 is a C6-C10 aryl group; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f , R e and R f are all C6-C8 straight chain alkyl groups; L3 and L4 are -OC(=O)-, wherein G1, G2, G3, G4, G5, G6, G7, L1 and L2 are as defined in formula (I).

[0197] Preferably, R4 is a C1-C12 alkyl group, R4, G5 or a portion of G5 together with the nitrogen to which it is attached forms a ring; R3 is a C1-C3 alkyl group; L3 and L4 are -OC(=O)-; L5 and L6 are both -C(=O)NR e Rf ,R e and R f are all C6-C8 linear alkyl groups, wherein the definitions of G1, G2, G3, G4, G5, G6, G7, L1, and L2 are as defined in formula (I).

[0198] Preferably, R4 is a C1-C3 alkyl group; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f ,R e and R f are all C6-C8 linear alkyl groups; L3 and L4 are -OC(=O)-, wherein the definitions of G1, G2, G3, G4, G5, G6, G7, L1, and L2 are as defined in formula (I).

[0199] Preferably, R4 is a C6-C10 aryl group; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f ,R e and R f are all C6-C8 linear alkyl groups; L3 and L4 are -OC(=O)-, wherein the definitions of G1, G2, G3, G4, G5, G6, G7, L1, and L2 are as defined in formula (I).

[0200] Preferably, R4 is a C1-C12 alkyl group, and R4, G5 or a part of G5 together with the nitrogen to which it is attached form a ring; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f ,R e and R f are all C6-C8 linear alkyl groups; L3 and L4 are -OC(=O)-, wherein the definitions of G1, G2, G3, G4, G5, G6, G7, L1, and L2 are as defined in formula (I).

[0201] Preferably, R3 and R4 are C1-C12 alkyl groups, and R3 and R4 together with the nitrogen to which they are attached form a ring; L5 and L6 are both -C(=O)NR e R f ,R e and R f are all C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-, wherein the definitions of G1, G2, G3, G4, G5, G6, G7 are as defined in formula (I).

[0202] Preferably, R4 is a C1-C3 alkyl group; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e Rf , R e and R f are all C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-, where the definitions of G1, G2, G3, G4, G5, G6, and G7 are as defined in formula (I).

[0203] Preferably, R4 is a C6-C10 aryl group; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f , R e and R f are all C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-, where the definitions of G1, G2, G3, G4, G5, G6, and G7 are as defined in formula (I).

[0204] Preferably, R4 is a C1-C12 alkyl group, and R4, G5 or a part of G5 together with the nitrogen to which it is attached form a ring; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f , R e and R f are all C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-, where the definitions of G1, G2, G3, G4, G5, G6, and G7 are as defined in formula (I).

[0205] Preferably, R4 is a C1-C3 alkyl group; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f , R e and R f are all C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-, where the definitions of G1, G2, G3, G4, G5, G6, and G7 are as defined in formula (I).

[0206] Preferably, R4 is a C6-C10 aryl group; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f , R e and R f are all C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-, where the definitions of G1, G2, G3, G4, G5, G6, and G7 are as defined in formula (I).

[0207] Preferably, R4 is a C1-C12 alkyl group, and R4, G5 or a part of G5 together with the nitrogen to which it is attached form a ring; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f , R e and R f are all C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-, where the definitions of G1, G2, G3, G4, G5, G6, and G7 are as defined in formula (I).

[0208] Preferably, R3 and R4 are C1-C12 alkyl groups, and R3, R4 together with the nitrogen to which they are attached form a ring; L5 and L6 are both -C(=O)NR e R f , R e and R f are all C6-C8 straight-chain alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-, where the definitions of G1, G2, G3, G4, G5, G6, and G7 are as defined in formula (I).

[0209] Preferably, R4 is a C1-C3 alkyl group; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f , R e and R f are all C6-C8 straight-chain alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-, where the definitions of G1, G2, G3, G4, G5, G6, and G7 are as defined in formula (I).

[0210] Preferably, R4 is a C6-C10 aryl group; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f , R e and R f are all C6-C8 straight-chain alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-, where the definitions of G1, G2, G3, G4, G5, G6, and G7 are as defined in formula (I).

[0211] Preferably, R4 is a C1-C12 alkyl group, and R4, G5 or a part of G5 together with the nitrogen to which it is attached form a ring; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f , R e and R fAll are straight-chain C6-C8 alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-, where the definitions of G1, G2, G3, G4, G5, G6, and G7 are as defined in formula (I).

[0212] Preferably, R4 is a C1-C3 alkyl group; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f R e and R f All are straight-chain C6-C8 alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-, where the definitions of G1, G2, G3, G4, G5, G6, and G7 are as defined in formula (I).

[0213] Preferably, R4 is a C6-C10 aryl group; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f R e and R f All are straight-chain C6-C8 alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-, where the definitions of G1, G2, G3, G4, G5, G6, and G7 are as defined in formula (I).

[0214] Preferably, R4 is a C1-C12 alkyl group, and R4, G5 or a part of G5 together with the nitrogen to which it is attached form a ring; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f R e and R f All are straight-chain C6-C8 alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-, where the definitions of G1, G2, G3, G4, G5, G6, and G7 are as defined in formula (I).

[0215] Preferably, R3 and R4 are C1-C12 alkyl groups, and R3, R4 together with the nitrogen to which they are attached form a ring; L5 and L6 are both -C(=O)NR e R f R e and R f All are branched-chain C6-C8 alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group, where the definitions of G1, G2, G3, G4, G6, and G7 are as defined in formula (I).

[0216] Preferably, R4 is a C1-C3 alkyl group; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f ,R e and R f are both C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group, wherein the definitions of G1, G2, G3, G4, G6, and G7 are as defined in formula (I).

[0217] Preferably, R4 is a C6-C10 aryl group; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f ,R e and R f are both C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group, wherein the definitions of G1, G2, G3, G4, G6, and G7 are as defined in formula (I).

[0218] Preferably, R4 is a C1-C12 alkyl group, and a part of R4 or G5 together with the nitrogen to which it is attached forms a ring; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f ,R e and R f are both C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-, wherein the definitions of G1, G2, G3, G4, G6, and G7 are as defined in formula (I).

[0219] Preferably, R4 is a C1-C3 alkyl group; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f ,R e and R f are both C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group, wherein the definitions of G1, G2, G3, G4, G6, and G7 are as defined in formula (I).

[0220] Preferably, R4 is a C6-C10 aryl group; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f ,R e and R fAll are C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group, wherein the definitions of G1, G2, G3, G4, G6, and G7 are as defined in formula (I).

[0221] Preferably, R4 is a C1-C12 alkyl group, and a part of R4 or G5 together with the nitrogen to which it is attached forms a ring; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f ,R e and R f All are C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group, wherein the definitions of G1, G2, G3, G4, G6, and G7 are as defined in formula (I).

[0222] Preferably, R3 and R4 are C1-C12 alkyl groups, and R3, R4 together with the nitrogen to which they are attached form a ring; L5 and L6 are both -C(=O)NR e R f ,R e and R f All are C6-C8 straight-chain alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group, wherein the definitions of G1, G2, G3, G4, G6, and G7 are as defined in formula (I).

[0223] Preferably, R4 is a C1-C3 alkyl group; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f ,R e and R f All are C6-C8 straight-chain alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group, wherein the definitions of G1, G2, G3, G4, G6, and G7 are as defined in formula (I).

[0224] Preferably, R4 is a C6-C10 aryl group; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f ,R e and R f All are C6-C8 straight-chain alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group, wherein the definitions of G1, G2, G3, G4, G6, and G7 are as defined in formula (I).

[0225] Preferably, R4 is a C1-C12 alkyl group, and a part of R4 or G5 together with the nitrogen to which it is attached forms a ring; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f R e and R f are all C6-C8 straight-chain alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group, wherein the definitions of G1, G2, G3, G4, G6, and G7 are as defined in formula (I).

[0226] Preferably, R4 is a C1-C3 alkyl group; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f R e and R f are all C6-C8 straight-chain alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group, wherein the definitions of G1, G2, G3, G4, G6, and G7 are as defined in formula (I).

[0227] Preferably, R4 is a C6-C10 aryl group; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f R e and R f are all C6-C8 straight-chain alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group, wherein the definitions of G1, G2, G3, G4, G6, and G7 are as defined in formula (I).

[0228] Preferably, R4 is a C1-C12 alkyl group, and a part of R4 or G5 together with the nitrogen to which it is attached forms a ring; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f R e and R f are all C6-C8 straight-chain alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group, wherein the definitions of G1, G2, G3, G4, G6, and G7 are as defined in formula (I).

[0229] Preferably, R3 and R4 are C1-C12 alkyl groups, and R3 and R4 together with the nitrogen to which they are attached form a ring; L5 and L6 are both -C(=O)NR e Rf and R e and R f are each a C6-C8 branched alkyl group; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups, where G1, G2, G3, and G4 are as defined in formula (I).

[0230] Preferably, R4 is a C1-C3 alkyl group; R3 is a C1-C3 alkyl group; L5 and L6 are each -C(=O)NR e R f and R e and R f are each a C6-C8 branched alkyl group; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups, where G1, G2, G3, and G4 are as defined in formula (I).

[0231] Preferably, R4 is a C6-C10 aryl group; R3 is a C1-C3 alkyl group; L5 and L6 are each -C(=O)NR e R f and R e and R f are each a C6-C8 branched alkyl group; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups, where G1, G2, G3, and G4 are as defined in formula (I).

[0232] Preferably, R4 is a C1-C12 alkyl group, and a part of R4 or G5 together with the nitrogen to which it is attached forms a ring; R3 is a C1-C3 alkyl group; L5 and L6 are each -C(=O)NR e R f and R e and R f are each a C6-C8 branched alkyl group; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G6 and G7 are unsubstituted C2-C4 alkylene groups, where G1, G2, G3, and G4 are as defined in formula (I).

[0233] Preferably, R4 is a C1-C3 alkyl group; R3 is G1L1G3L3L5; L5 and L6 are each -C(=O)NR e R f and R e and R fAll are C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups, wherein the definitions of G1, G2, G3, and G4 are as defined in formula (I).

[0234] Preferably, R4 is a C6-C10 aryl group; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f ,R e and R f All are C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups, wherein the definitions of G1, G2, G3, and G4 are as defined in formula (I).

[0235] Preferably, R4 is a C1-C12 alkyl group, and a part of R4 or G5 together with the nitrogen to which it is attached forms a ring; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f ,R e and R f All are C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups, wherein the definitions of G1, G2, G3, and G4 are as defined in formula (I).

[0236] Preferably, R3 and R4 are C1-C12 alkyl groups, and R3, R4 together with the nitrogen to which they are attached form a ring; L5 and L6 are both -C(=O)NR e R f ,R e and R f All are C6-C8 straight-chain alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups, wherein the definitions of G1, G2, G3, and G4 are as defined in formula (I).

[0237] Preferably, R4 is a C1-C3 alkyl group; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f ,R e and R fAll are straight-chain C6-C8 alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups, wherein the definitions of G1, G2, G3, and G4 are as defined in formula (I).

[0238] Preferably, R4 is a C6-C10 aryl group; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f ,R e and R f All are straight-chain C6-C8 alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups, wherein the definitions of G1, G2, G3, and G4 are as defined in formula (I).

[0239] Preferably, R4 is a C1-C12 alkyl group, a part of R4 or G5 together with the nitrogen to which it is attached forms a ring; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f ,R e and R f All are straight-chain C6-C8 alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups, wherein the definitions of G1, G2, G3, and G4 are as defined in formula (I).

[0240] Preferably, R4 is a C1-C3 alkyl group; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f ,R e and R f All are straight-chain C6-C8 alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups, wherein the definitions of G1, G2, G3, and G4 are as defined in formula (I).

[0241] Preferably, R4 is a C6-C10 aryl group; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f ,R e and R fAll are straight-chain C6-C8 alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups, where the definitions of G1, G2, G3, and G4 are as defined in formula (I).

[0242] Preferably, R4 is a C1-C12 alkyl group, and a part of R4 or G5 together with the nitrogen to which it is attached forms a ring; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f ,R e and R f All are straight-chain C6-C8 alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups, where the definitions of G1, G2, G3, and G4 are as defined in formula (I).

[0243] Preferably, R3 and R4 are C1-C12 alkyl groups, and R3, R4 together with the nitrogen to which they are attached form a ring; L5 and L6 are both -C(=O)NR e R f ,R e and R f All are branched-chain C6-C8 alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups; G3 and G4 are unsubstituted C2-C4 alkylene groups, where the definitions of G1 and G2 are as defined in formula (I).

[0244] Preferably, R4 is a C1-C3 alkyl group; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f ,R e and R f All are branched-chain C6-C8 alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups; G3 and G4 are unsubstituted C2-C4 alkylene groups, where the definitions of G1 and G2 are as defined in formula (I).

[0245] Preferably, R4 is a C6-C10 aryl group; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f ,R e and R fAll are C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups; G3 and G4 are unsubstituted C2-C4 alkylene groups, wherein the definitions of G1 and G2 are as defined in formula (I).

[0246] Preferably, R4 is a C1-C12 alkyl group, and a part of R4 or G5 together with the nitrogen to which it is attached forms a ring; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f ,R e and R f All are C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G6 and G7 are unsubstituted C2-C4 alkylene groups; G3 and G4 are unsubstituted C2-C4 alkylene groups, wherein the definitions of G1 and G2 are as defined in formula (I).

[0247] Preferably, R4 is a C1-C3 alkyl group; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f ,R e and R f All are C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups; G3 and G4 are unsubstituted C2-C4 alkylene groups, wherein the definitions of G1 and G2 are as defined in formula (I).

[0248] Preferably, R4 is a C6-C10 aryl group; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f ,R e and R f All are C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups; G3 and G4 are unsubstituted C2-C4 alkylene groups, wherein the definitions of G1 and G2 are as defined in formula (I).

[0249] Preferably, R4 is a C1-C12 alkyl group, and a part of R4 or G5 together with the nitrogen to which it is attached forms a ring; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f ,R e and Rf All are C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups; G3 and G4 are unsubstituted C2-C4 alkylene groups, wherein the definitions of G1 and G2 are as defined in formula (I).

[0250] Preferably, R3 and R4 are C1-C12 alkyl groups, and R3, R4 together with the nitrogen atom to which they are attached form a ring; L5 and L6 are both -C(=O)NR e R f R e and R f All are C6-C8 straight-chain alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups; G3 and G4 are unsubstituted C2-C4 alkylene groups, wherein the definitions of G1 and G2 are as defined in formula (I).

[0251] Preferably, R4 is a C1-C3 alkyl group; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f R e and R f All are C6-C8 straight-chain alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups; G3 and G4 are unsubstituted C2-C4 alkylene groups, wherein the definitions of G1 and G2 are as defined in formula (I).

[0252] Preferably, R4 is a C6-C10 aryl group; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f R e and R f All are C6-C8 straight-chain alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups; G3 and G4 are unsubstituted C2-C4 alkylene groups, wherein the definitions of G1 and G2 are as defined in formula (I).

[0253] Preferably, R4 is a C1-C12 alkyl group, and a part of R4 or G5 together with the nitrogen atom to which they are attached forms a ring; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f R eand R f Both are C6-C8 straight-chain alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups; G3 and G4 are unsubstituted C2-C4 alkylene groups, wherein the definitions of G1 and G2 are as defined in formula (I).

[0254] Preferably, R4 is a C1-C3 alkyl group; R3 is G1L1G3L3L5; both L5 and L6 are -C(=O)NR e R f R e and R f Both are C6-C8 straight-chain alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups; G3 and G4 are unsubstituted C2-C4 alkylene groups, wherein the definitions of G1 and G2 are as defined in formula (I).

[0255] Preferably, R4 is a C6-C10 aryl group; R3 is G1L1G3L3L5; both L5 and L6 are -C(=O)NR e R f R e and R f Both are C6-C8 straight-chain alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups; G3 and G4 are unsubstituted C2-C4 alkylene groups, wherein the definitions of G1 and G2 are as defined in formula (I).

[0256] Preferably, R4 is a C1-C12 alkyl group, and a part of R4 or G5 together with the nitrogen to which it is attached forms a ring; R3 is G1L1G3L3L5; both L5 and L6 are -C(=O)NR e R f R e and R f Both are C6-C8 straight-chain alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups; G3 and G4 are unsubstituted C2-C4 alkylene groups, wherein the definitions of G1 and G2 are as defined in formula (I).

[0257] Preferably, R3 and R4 are C1-C12 alkyl groups, and R3, R4 together with the nitrogen to which they are attached form a ring; both L5 and L6 are -C(=O)NR e R f, R e and R f are each a C6-C8 branched alkyl group; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups; G3 and G4 are unsubstituted C2-C4 alkylene groups; G1 and G2 are unsubstituted C2-C4 alkylene groups.

[0258] Preferably, R4 is a C1-C3 alkyl group; R3 is a C1-C3 alkyl group; L5 and L6 are each -C(=O)NR e R f , R e and R f are each a C6-C8 branched alkyl group; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups; G3 and G4 are unsubstituted C2-C4 alkylene groups; G1 and G2 are unsubstituted C2-C4 alkylene groups.

[0259] Preferably, R4 is a C6-C10 aryl group; R3 is a C1-C3 alkyl group; L5 and L6 are each -C(=O)NR e R f , R e and R f are each a C6-C8 branched alkyl group; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups; G3 and G4 are unsubstituted C2-C4 alkylene groups; G1 and G2 are unsubstituted C2-C4 alkylene groups.

[0260] Preferably, R4 is a C1-C12 alkyl group, a part of R4 or G5 together with the nitrogen to which it is attached forms a ring; R3 is a C1-C3 alkyl group; L5 and L6 are each -C(=O)NR e R f , R e and R f are each a C6-C8 branched alkyl group; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G6 and G7 are unsubstituted C2-C4 alkylene groups; G3 and G4 are unsubstituted C2-C4 alkylene groups; G1 and G2 are unsubstituted C2-C4 alkylene groups.

[0261] Preferably, R4 is a C1-C3 alkyl group; R3 is G1L1G3L3L5; L5 and L6 are each -C(=O)NR e R f , R e and Rf All are C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups; G3 and G4 are unsubstituted C2-C4 alkylene groups; G1 and G2 are unsubstituted C2-C4 alkylene groups.

[0262] Preferably, R4 is a C6-C10 aryl group; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f R e and R f All are C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups; G3 and G4 are unsubstituted C2-C4 alkylene groups; G1 and G2 are unsubstituted C2-C4 alkylene groups.

[0263] Preferably, R4 is a C1-C12 alkyl group, and a part of R4 or G5 together with the nitrogen to which it is attached forms a ring; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f R e and R f All are C6-C8 branched alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups; G3 and G4 are unsubstituted C2-C4 alkylene groups; G1 and G2 are unsubstituted C2-C4 alkylene groups.

[0264] Preferably, R3 and R4 are C1-C12 alkyl groups, and R3, R4 together with the nitrogen to which they are attached form a ring; L5 and L6 are both -C(=O)NR e R f R e and R f All are C6-C8 straight-chain alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups; G3 and G4 are unsubstituted C2-C4 alkylene groups; G1 and G2 are unsubstituted C2-C4 alkylene groups, R4 is a C1-C3 alkyl group; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f R e and R fAll are straight-chain C6-C8 alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups; G3 and G4 are unsubstituted C2-C4 alkylene groups; G1 and G2 are unsubstituted C2-C4 alkylene groups.

[0265] Preferably, R4 is a C6-C10 aryl group; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f R e and R f All are straight-chain C6-C8 alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups; G3 and G4 are unsubstituted C2-C4 alkylene groups; G1 and G2 are unsubstituted C2-C4 alkylene groups.

[0266] Preferably, R4 is a C1-C12 alkyl group, and a part of R4 or G5 together with the nitrogen to which it is attached forms a ring; R3 is a C1-C3 alkyl group; L5 and L6 are both -C(=O)NR e R f R e and R f All are straight-chain C6-C8 alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups; G3 and G4 are unsubstituted C2-C4 alkylene groups; G1 and G2 are unsubstituted C2-C4 alkylene groups.

[0267] Preferably, R4 is a C1-C3 alkyl group; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f R e and R f All are straight-chain C6-C8 alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups; G3 and G4 are unsubstituted C2-C4 alkylene groups, wherein; G1 and G2 are unsubstituted C2-C4 alkylene groups.

[0268] Preferably, R4 is a C6-C10 aryl group; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f R e and Rf All are C6-C8 straight-chain alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups; G3 and G4 are unsubstituted C2-C4 alkylene groups; G1 and G2 are unsubstituted C2-C4 alkylene groups.

[0269] Preferably, R4 is a C1-C12 alkyl group, and a part of R4 or G5 together with the nitrogen to which it is attached forms a ring; R3 is G1L1G3L3L5; L5 and L6 are both -C(=O)NR e R f R e and R f All are C6-C8 straight-chain alkyl groups; L3 and L4 are -OC(=O)-; L1 and L2 are -C(=O)O-; G5 is an unsubstituted C2-C5 alkylene group; G6 and G7 are unsubstituted C2-C4 alkylene groups; G3 and G4 are unsubstituted C2-C4 alkylene groups; G1 and G2 are unsubstituted C2-C4 alkylene groups.

[0270] More preferably, the compound has the structure shown in formula (I-A-1):

[0271]

[0272] wherein m and n are each independently 1 or 3, and the definitions of R e and R f are as defined in formula (I).

[0273] Or preferably, the compound has the structure shown in formula (I-A-2):

[0274]

[0275]

[0276] wherein m and n are each independently 1 or 3, R5 and R6 are optionally substituted C1-C12 alkyl groups, optionally substituted C2-C12 alkenyl groups, optionally substituted C2-C12 alkynyl groups, optionally substituted C6-C10 aryl groups or 5- to 10-membered heteroaryl groups; or R5, R6 together with the nitrogen to which they are attached form a ring, and the definitions of G5, R e and R f are as defined in formula (I).

[0277] Or preferably, the compound has the structure shown in formula (I-B-1):

[0278]

[0279] Among them, m and n are each independently 1 or 3, R e and R f are defined as defined in formula (I).

[0280] More preferably, R e and / or R f has one of the following structures:

[0281]

[0282] This application provides the following compounds or pharmaceutically acceptable salts, prodrugs or stereoisomers thereof:

[0283] Table 1 Representative Compounds

[0284]

[0285]

[0286]

[0287]

[0288]

[0289]

[0290]

[0291]

[0292]

[0293]

[0294]

[0295]

[0296]

[0297]

[0298]

[0299]

[0300]

[0301]

[0302]

[0303]

[0304] The present application further provides a composition, comprising a therapeutic or prophylactic agent; and a carrier for delivering the therapeutic or prophylactic agent, wherein the carrier comprises an ionizable lipid, and the ionizable lipid comprises one or more of the ionizable lipid compounds provided in any of the foregoing applications of the present application, or a pharmaceutically acceptable salt thereof.

[0305] In the composition of the present application, the therapeutic or prophylactic agent may be a nucleic acid molecule, a small molecule compound, a polypeptide or a protein, or a mixture of two or more thereof.

[0306] In a specific embodiment, the nucleic acid molecule is selected from single-stranded DNA, double-stranded DNA, short isoforms, agomir, antagomir, antisense molecules, small interfering RNA (siRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), Dicer substrate RNA (dsRNA), small hairpin RNA (shRNA), transfer RNA (tRNA), messenger RNA (mRNA), and other forms of RNA molecules known in the art, or nucleic acid mimetics such as locked nucleic acid (LNA), peptide nucleic acid (PNA), and morpholino oligonucleotides.

[0307] In a specific embodiment, the therapeutic or prophylactic agent comprises at least one mRNA encoding an antigen or a fragment or epitope thereof, preferably, the mRNA is a monocistronic mRNA or a polycistronic mRNA.

[0308] In a specific embodiment, the antigen is a pathogenic antigen.

[0309] In a specific embodiment, the mRNA comprises one or more functional nucleotide analogs, preferably, the functional nucleotide analogs are selected from one or more of pseudouridine, 1-methyl-pseudouridine, or 5-methylcytosine.

[0310] In a specific embodiment, the small molecule compound is selected from one or more of anti-tumor drugs, anti-infective drugs, local anesthetics, antidepressants, anti-convulsants, antibiotics / antibacterials, anti-fungal drugs, anti-parasitic drugs, hormones, hormone antagonists, immunomodulators, neurotransmitter antagonists, anti-glaucoma agents, anesthetics, or imaging agents.

[0311] As described above, in the composition provided by the present application, the amounts of the carrier and the therapeutic or prophylactic agent are not limited in the present application. In a specific embodiment, the mass ratio of the carrier to the therapeutic or prophylactic agent is 5:1 to 50:1. For example, it can be 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, 25:1, 30:1, 35:1, 40:1, 41:1, 42:1, 43:1, 44:1, 45:1, 46:1, 47:1, 48:1, 49:1.

[0312] In a specific embodiment, the composition of the present application is a nanoparticle preparation, and the average size of the nanoparticle preparation is 10 to 500 nm. For example, it can be 20 nm, 30 nm, 40 nm, 50 nm, 60 nm, 70 nm, 80 nm, 90 nm, 100 nm, 150 nm, 200 nm, 250 nm, 300 nm, 350 nm, 400 nm, 450 nm.

[0313] In a specific embodiment, the pKa of the nanoparticle preparation of the present application is 4.5 to 8.5. For example, it can be 4.6, 4.7, 4.8, 4.9, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6, 6.5, 7, 7.5, 8, 8.1, 8.2, 8.3, 8.4.

[0314] As described above, in the composition provided by the present application, the carrier further comprises one or more neutral lipids. Preferably, the neutral lipid is selected from one or more of phosphatidylcholine, phosphatidylethanolamine, sphingomyelin, ceramide, sterol and its derivatives.

[0315] In a specific embodiment, the molar ratio of the ionizable lipid to the neutral lipid is in the range of about 100:1 to about 5:1. For example, it can be 90:1, 85:1, 80:1, 75:1, 70:1, 65:1, 60:1, 55:1, 50:1, 45:1, 40:1, 35:1, 30:1, 25:1, 20:1, 15:1, 10:1.

[0316] In a specific embodiment, the steroid is selected from one or more of cholesterol, non-sterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatine, tomatidine, ursolic acid, α-tocopherol, corticosteroid.

[0317] In a specific embodiment, the molar ratio of the ionizable lipid to the steroid ranges from about 2:1 to about 4:1, and for example, it can be 2.1:1, 2.2:1, 2.3:1, 2.4:1, 2.5:1, 2.6:1, 2.7:1, 2.8:1, 2.9:1, 3:1, 3.1:1, 3.2:1, 3.3:1, 3.4:1, 3.5:1, 3.6:1, 3.7:1, 3.8:1, 3.9:1.

[0318] As described above, the composition provided by the present application, wherein the composition carrier further comprises one or more lipids capable of binding to the polymer. Preferably, the lipid capable of binding to the polymer is one or more selected from PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, and PEG-modified dialkylglycerol.

[0319] In a specific embodiment, the molar ratio of the ionizable lipid to the lipid capable of binding to the polymer ranges from about 100:1 to about 20:1, and for example, it can be 95:1, 90:1, 85:1, 80:1, 75:1, 70:1, 65:1, 60:1, 55:1, 50:1, 45:1, 40:1, 35:1, 30:1, 25:1.

[0320] In a specific embodiment, in the composition of the present application, the carrier further comprises neutral lipid, structural lipid, and polymer-conjugated lipid, and the molar ratio of the ionizable lipid, the neutral lipid, the steroid lipid, and the polymer-conjugated lipid is (15-70):(1-15):(15-35):(0-3).

[0321] In a specific embodiment, in the composition of the present application, the carrier further comprises neutral lipid and structural lipid, and the molar ratio of the ionizable lipid, the neutral lipid, and the steroid lipid is (15-70):(1-15):(15-35).

[0322] In a specific embodiment, the composition further comprises a pharmaceutically acceptable excipient.

[0323] In a specific embodiment, the excipient includes a pharmaceutically acceptable diluent.

[0324] The present application further provides the use of the compound represented by formula (I) provided by any one of the above in the present application, or a pharmaceutically acceptable salt, prodrug, or stereoisomer thereof in the preparation of a drug.

[0325] The present application further provides the use of the composition provided by any one of the above in the present application in the preparation of a drug.

[0326] In a specific embodiment, the drug is any one selected from gene drugs, nucleic acid vaccines, small molecule drugs, polypeptides or protein drugs.

[0328] The following further clarifies the present application in conjunction with specific embodiments, but these examples are only used to illustrate the present application and not to limit the scope of the present application. The implementation conditions adopted in the examples can be further adjusted according to different requirements of specific use, and the implementation conditions not specified are conventional conditions in the industry. The technical features involved in each embodiment of the present application can be combined with each other as long as they do not conflict with each other.

[0329] In the specific examples of the present application, the raw materials used can all be obtained commercially.

[0330] Example 1

[0331] Synthesis route of compound A1-H226

[0332]

[0333] Step 1: Synthesis of intermediate H226

[0334] To a dichloromethane (200 mL) solution of diethylamine (10 g, 54 mmol, 1 eq) and succinic anhydride (6.49 g, 65 mmol, 1.2 eq), (15 mL, 108 mmol, 2 eq) was added. The reaction mixture was stirred at room temperature overnight, adjusted to pH neutral with 1 M hydrochloric acid solution, and washed with brine. The combined organic layers were dried over MgSO4 and the solvent was removed in vacuo to obtain the crude product H226-1. In a 500 mL round-bottom flask containing the crude product H226-1, hydroxyethyl acrylate (7.52 g, 65 mmol, 1.2 eq), DMAP (1.34 g, 11 mmol, 0.2 eq) and dichloromethane solvent (300 mL) were added. After the mixture was stirred at room temperature for 5 minutes, DCC (13.4 g, 65 mmol, 1.2 eq) dissolved in dichloromethane was added dropwise. The reaction mixture was stirred at room temperature overnight, and TLC showed that compound H226-1 had completely disappeared. It was washed with brine, the combined organic layers were dried over MgSO4 and the solvent was removed in vacuo to obtain the crude product. The crude product was purified by column chromatography (eluent: PE / EA = 5 / 1), and the pure product fraction was evaporated to obtain a slightly yellow oily compound H226 (8.06 g, 39%). 11H NMR (400 MHz, CDCl3) δ = 6.44 (ddd, J = 17.3, 1.3, 0.8, 1H), 6.15 (ddd, J = 17.3, 10.4, 0.7, 1H), 5.86 (ddd, J = 10.4, 1.3, 0.8, 1H), 4.39 - 4.29 (m, 4H), 3.37 - 3.11 (m, 4H), 2.66 (dq, J = 12.8, 6.6, 4H), 1.65 - 1.42 (m, 4H), 1.33 - 1.26 (m, 12H), 0.89 (dt, J = 10.6, 6.7, 6H). MS m / z (ESI): ***[M+H] +

[0335] The following compounds were prepared in a similar manner to intermediate H226 using the corresponding starting materials.

[0336] Table 2 Representative intermediates and structure confirmation data

[0337]

[0338]

[0339]

[0340] Step 2: Synthesis of compound A1-H226

[0341] At 70 °C, N,N-dimethylethylenediamine (29 μL, 0.26 mmol, 1 eq) and compound H226 (300 μL, 0.78 mmol, 3 eq) were mixed and stirred for 48 h. The mixture was purified by column chromatography (silica gel column, eluent: dichloromethane solution containing 0 - 10% methanol (volume percentage)) to obtain compound A1-H226 (120 mg, 54%), which was a slightly yellow oil. 1 1H NMR (400 MHz, CDCl3). MS m / z (ESI): 855.6 [M+H] + 。

[0342] The following compounds were prepared in a similar manner to compound A1-H226 using the corresponding starting materials.

[0343] Table 3 Representative compounds and structure confirmation data

[0344]

[0345]

[0346]

[0347]

[0348]

[0349]

[0350]

[0351]

[0352]

[0353]

[0354]

[0355]

[0356]

[0357]

[0358]

[0359]

[0360] Example 2

[0361] Preparation of nanoparticle composition

[0362] 75 ionizable lipid compounds prepared in Example 1 were respectively dissolved in absolute ethanol with cholesterol (Avitide (Shanghai) Pharmaceutical Technology Co., Ltd.), DSPC alcohol (Avitide (Shanghai) Pharmaceutical Technology Co., Ltd.) and DMG-PEG2000 (Avitide (Shanghai) Pharmaceutical Technology Co., Ltd.) at a molar ratio of 66.67:25.67:6.67:1.00. Lipid nanoparticles (LNPs) were prepared at a weight ratio of approximately 10:1 of ionizable lipid to firefly luciferase (Fluc) mRNA. Briefly, the mRNA was diluted in 25 mM sodium acetate solution (pH 5.2), and the ethanol solution of the lipid was mixed with the aqueous mRNA solution at a ratio of approximately 1:3 (volume / volume) using a syringe pump, with a total flow rate of 12 mL / min. The ethanol was removed by dialysis to displace the LNPs into PBS buffer. Finally, the lipid nanoparticles were filtered through a sterile filter with a 0.2 μm pore size to obtain an LNP preparation (LNP-mFluc) encapsulating firefly luciferase mRNA.

[0363] Example 3

[0364] In vivo evaluation of luciferase mRNA using the lipid nanoparticle composition

[0365] The luciferase mRNA in vivo evaluation was performed on the lipid nanoparticle composition (LNP-mFluc) prepared in Example 2. FLuc mRNA from Shanghai Hexincheng Biotechnology will express luciferase protein, which was initially isolated from fireflies. Fluc is commonly used in mammalian cell cultures to measure gene expression and cell viability. It emits bioluminescence in the presence of the substrate luciferin. The study was conducted on 5-6-week-old female Balb / c mice (Shanghai Slack Experimental Animal Co., Ltd.) according to the guidelines established by the Animal Care Committee (ACC) and the Canadian Council on Animal Care (CCAC). After tail vein injection of 5 μg luciferase mRNA and 50 μg ionizable lipid nanoparticles, 100 μL of 30 mg / mL D-luciferin potassium salt (Adamas Reagent Co., Ltd.) was intraperitoneally injected into the mice 6 h after administration. The mice were imaged using a live imaging system (PerkinElmer) 10 minutes later. The in vivo delivery efficiency and location of 75 LNP-mFluc are as Figure 1 and Figure 2 shown, and the fluorescence intensity in the spleen is shown in Table 4.

[0366] Table 4 Fluorescence intensity of LNP-mFluc prepared from 75 compounds in the spleen

[0367]

[0368]

[0369] Example 4

[0370] Determination of the efficacy (formulation optimization) of lipid nanoparticle formulations containing A1-H226x ionizable lipid using a rodent model of in vivo luciferase mRNA expression

[0371] A. To test the effect of changes in DSPC content and cholesterol content on delivery efficiency, lipid nanoparticles were prepared by the preparation method described in Example 2 with the molar ratio scheme shown in Table 5. After tail vein injection of 5 μg luciferase mRNA and 50 μg A1-H226x ionizable lipid nanoparticles according to the experimental method described in Example 4, the test results are shown in Figure 3 .

[0372] Table 5 Experiment on the effect of changes in DSPC / cholesterol content on expression efficiency (molar ratio)

[0373]

[0374]

[0375] B. Test the effects of the content of DEM-PEG2000 and the content of ionizable lipids on the delivery efficiency. Prepare lipid nanoparticles according to the preparation method described in Example 2 with the molar ratio scheme in Table 6. Inject 5 μg luciferase mRNA and 50 μg A1-H226x ionizable lipid nanoparticles via the tail vein according to the experimental method described in Example 4. The test results are shown in Figure 4 .

[0376] Table 6 Experiment on the effects of changes in the content of DMG-PEG2000 and ionizable lipids on the expression efficiency (molar ratio)

[0377]

[0378] C. Test the effects of phospholipids on the delivery efficiency. Prepare lipid nanoparticles according to the preparation method described in Example 2 with the molar ratio scheme in Table 7. Inject 5 μg luciferase mRNA and 50 μg A1-H226x ionizable lipid nanoparticles via the tail vein according to the experimental method described in Example 4. The test results are shown in Figure 5 .

[0379] Table 7 Experiment on the effects of different phospholipids on the expression efficiency (molar ratio)

[0380]

[0381] Example 5

[0382] Immunoassay

[0383] The study was conducted on female C57BL / 6 mice (Shanghai SIPPR-BK Laboratory Animal Co., Ltd.) at 6-8 weeks of age. The mice were subcutaneously inoculated with 1×10 ^6 B16F10-OVA cells on day 0. Prepare A1-H226x / OVA lipid nanoparticle composition and A4-H228 / OVA lipid nanoparticle composition according to the preparation method described in Example 2. Inject 10 μg A1-H226x / OVA or A4-H228 / OVA or do not treat (UT) via the tail vein on days 4, 7, and 10 respectively. After euthanizing the mice on day 14, collect blood, spleen, and tumor tissues.

[0384] A. For blood samples, after collection, they are placed at room temperature for 0.5 - 1 hour, centrifuged at 2000g for 20 minutes, and then the supernatant is separated, which is serum. The serum is stored in a -80°C refrigerator for subsequent determination of IgG antibody levels. The IgG antibody level is determined by ELISA method. Specifically: 100 μL per well of 10 μg / mL OVA peptide (GenScript Biotech Corporation) is used to coat the ELISA plate at 4°C overnight. Wash the plate 3 times with PBS (PBST) containing 0.5% Tween - 20. Then block it with 2% BSA at room temperature for 2 hours. The serum sample is diluted 100 - fold as the first dilution point, and then serially diluted 2 - fold until the 10th dilution point (diluted 51200 - fold). 100 μL per well of the serum sample at each dilution point is added and incubated at 37°C for 2 hours. After washing 3 times with PBST, 100 μL per well of HRP - anti - IgG antibody diluted 2000 - fold is added and incubated at 37°C in the dark for 1 hour. After washing 3 times with PBST, 100 μL of TMB is added for color development. Finally, read the absorbance at 450 nm. The measurement results are shown in Figure 6 。

[0385] B. For spleens, use the rubber stopper part of a 2 - mL syringe to grind them into single - cell homogenates, filter through a 70 - μM cell strainer, add 6 - fold volume of red blood cell lysis buffer (Sangon Biotech (Shanghai) Co., Ltd.), incubate on ice for 5 minutes, and then add an equal volume of PBS for neutralization. After centrifuging at 400g for 5 minutes, remove the supernatant and resuspend in 1 mL of PBS on ice for later use.

[0386] C. For tumor tissues, grind the collected tumor tissues with the rubber stopper part of a 2 - mL syringe and filter through a 70 - μM cell strainer. Add 3.5 mL of universal tissue lysis buffer (Ruidewo Life Science Co., Ltd.) and incubate with shaking at 37°C for 20 minutes. After centrifuging at 400g for 5 minutes, remove the supernatant, add 6 mL of red blood cell lysis buffer (Sangon Biotech (Shanghai) Co., Ltd.), incubate on ice for 5 minutes, and then add an equal volume of PBS for neutralization. After centrifuging at 400g for 5 minutes, remove the supernatant and resuspend in 1 mL of PBS on ice for later use.

[0387] D. For immune cell phenotyping, take 2×10 ^6 single - cells from spleens and 2×10 ^6 single - cells from tumor tissues and stain them with the fluorescent - labeled antibodies listed in Table 8 in the dark at 4°C for 1 hour. Then wash the cells with wash buffer (Thermo Fisher) and analyze them on a flow cytometer (Beckman). The immune cell phenotyping results in the tumor tissues of mice treated and untreated with the A1 - H226x / OVA lipid nanoparticle composition are shown in Figure 7 ,and the antigen - specific T - cell analysis results are shown in Figure 8 。The T - cell analysis results in the tumor tissues of mice treated and untreated with the A4 - H228 / OVA lipid nanoparticle composition are shown inFigure 9 The analysis results of T cells in splenocytes are shown in Figure 10 .

[0388] Table 8 Flow cytometry antibodies for immune cell typing

[0389]

[0390] E. For the analysis of IFNγ cytokine, a mouse IFN-γ pre-coated ELISPOT kit (BioTeke Corporation) was used for detection. 2 x 10 ^5 spleen single cells were added to the pre-coated 96-well plate. After stimulation with 2 μg / mL OVA peptide (GenScript Biotech Corporation) for 21 hours, the experiment was carried out according to the kit instructions. After air drying, the number of spots was counted on a fluorescence enzyme-linked immunosorbent analyzer (Beijing Antai Yongxin Medical Technology Co., Ltd.). The results are shown in Table 9 and Table 10.

[0391] Table 9 Number of IFN-γ test spots in A1-H226x / OVA treatment and control groups

[0392]

[0393]

[0394] Table 10 Number of IFN-γ test spots in A4-H228 / OVA treatment and control groups

[0395]

[0396] Example 6

[0397] Tumor treatment evaluation:

[0398] Studies were carried out on female Balb / c (Shanghai SIPPR-BK Laboratory Animal Co., Ltd.) mice at 6-8 weeks of age. Each mouse was subcutaneously inoculated with 5 x 10 ^5 CT26-Luc cells on day 0. The A4-H228 / gp70 lipid nanoparticle composition was prepared according to the method described in Example 2, and 20 μg of A4-H228 / gp70 was injected via the tail vein on days 7, 14, and 21, respectively, without treatment. The tumor size was measured regularly, and the mice were euthanized when the tumor grew to 2000 mm 3 . The tumor volume calculation formula: tumor volume (mm 3 ) = length (mm) × width ^2 (mm) / 2. The tumor growth curve is shown in Figure 11 .

[0399] Experiments have proven that the ionizable lipid compounds of the present application can deliver nucleic acid molecules, small molecule compounds, polypeptides, proteins, etc. The carrier prepared using the ionizable lipid compounds of the present application can successfully transport nucleic acid molecules into cells for expression.

[0400] It should be noted that although the technical solutions of the present application are introduced with specific examples, those skilled in the art can understand that the present application should not be limited thereto. The above has described the embodiments of the present application. The above description is exemplary and not exhaustive, and is also not limited to the disclosed embodiments. Many modifications and variations are obvious to those of ordinary skill in the art without departing from the scope and spirit of the described embodiments. The selection of the terms used in the present application is intended to best explain the principles of the embodiments, practical applications, or improvements to the technology in the market, or to enable other ordinary skilled in the art in the technical field to understand the disclosed embodiments of the present application.

Claims

1. A compound of formula (I): or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, wherein: each of G1 and G2 is independently an optionally substituted C2-C24 straight-chain alkylene group; L1 and L2 are each independently -OC(=O)-, -C(=O)O-, -OC(=O)O-, -C(=O)-, -O-, -S(O) x -, -SS-, -C(=O)S-, -SC(=O)-, -NR a C(=O)-, -C(=O)NR b -, -NR a C(=O)NR b -, -OC(=O)NR b -, -NR a C(=O)O-, -SC(=S)-, -C(=S)S-, -C(=S)-, -CH(OH)-, -P(=O)(OR b )O-, -(C6-C10 arylene)- or -(6- to 10-membered heteroarylene)-; R a and R b each independently is H, optionally substituted C1-C12 alkyl or optionally substituted C1-C12 alkenyl; each of G3 and G4 is independently an optionally substituted C2-C24 straight-chain alkylene group; L3 and L4 are each independently -OC(=O)-, -C(=O)O-, -OC(=O)O-, -C(=O)-, -O-, -S(O) x -, -SS-, -C(=O)S-, -SC(=O)-, -NR c C(=O)-, -C(=O)NR c -, -NR c C(=O)NR d -, -OC(=O)NR c -, -NR c C(=O)O-, -SC(=S)-, -C(=S)S-, -C(=S)-, -CH(OH)-, -P(=O)(OR c )O-, -(C6-C10 arylene)- or -(6- to 10-membered heteroarylene)-; R c and R d each independently is H, optionally substituted C1-C12 alkyl or optionally substituted C1-C12 alkenyl; x is 0, 1 or 2; each of G6 and G7 is independently an optionally substituted C2-C24 straight-chain alkylene group; Each of said L5 and L6 is independently -OC(=O)R e 、-C(=O)OR e 、-OC(=O)OR e 、-C(=O)R e 、-OR e 、-S(O) x R e 、-S-SR e 、-C(=O)SR e 、-SC(=O)R e 、-NR e C(=O)R f 、-C(=O)NR e R f 、-NR a C(=O)NR e R f 、-OC(=O)NR e R f 、-NR e C(=O)OR f 、-SC(=S)R e 、-C(=S)SR e 、-C(=S)R e 、-CH(OH)R e 、-P(=O)(OR e )(OR f )、-(C6-C10 arylene)-R e 、-(6- to 10-membered heteroarylene)-R e or R e ; R e and R f each independently is H, optionally substituted C1-C12 alkyl or optionally substituted C1-C12 alkenyl; G5 is an optionally substituted C1-C12 alkylene group; R3 is an optionally substituted C1-C12 alkyl group, an optionally substituted C2-C12 alkenyl group, an optionally substituted C2-C12 alkynyl group, an optionally substituted C6-C10 aryl group or a 5- to 10-membered heteroaryl group; or R3 and R4 together with the nitrogen to which they are attached form a ring; or R3 is G1L1G3L3L5; R4 is an optionally substituted C1-C12 alkyl group, an optionally substituted C2-C12 alkenyl group, an optionally substituted C2-C12 alkynyl group, an optionally substituted C6-C10 aryl group or a 5- to 10-membered heteroaryl group; or R4, G5 or a part of G5 together with the nitrogen to which they are attached form a ring; or R4 is G1L1G3L3L5.

2. The compound according to claim 1, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, characterized in that, The compound has the structure shown in formula (I-A): wherein, G1, G2, G3, G4, G5, G6, G7, L3, L4, L5, L6, R3 and R4 are as defined in claim 1.

3. The compound according to claim 1, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, characterized in that, The compound has the structure shown in formula (I-B): wherein, the definitions of G1, G2, G3, G4, G5, G6, G7, L1, L2, L3, L4, L5, L6 and R3 are as defined in claim 1.

4. The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, characterized in that, wherein G1 and G2 are unsubstituted C2-C4 alkylene groups.

5. The compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, characterized in that, wherein G3 and G4 are unsubstituted C2-C4 alkylene groups; or wherein G6 and G7 are unsubstituted C2-C4 alkylene groups.

6. The compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, characterized in that, wherein L1 and L2 are -C(=O)O-.

7. A compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, characterized in that, wherein L3 and L4 are -OC(=O)-.

8. A compound according to any one of claims 1 to 7, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, characterized in that, Both L5 and L6 are -C(=O)NR e R f .

9. The compound according to claim 2, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, characterized in that, The compound has the structure shown in formula (I-A-1): wherein m and n are each independently 1 or 3, R e and R f are as defined in claim 1.

10. The compound according to claim 2, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, characterized in that, The compound has the structure shown in formula (I-A-2): wherein m and n are each independently 1 or 3, R5 and R6 are each independently an optionally substituted C1-C12 alkyl group, an optionally substituted C2-C12 alkenyl group, an optionally substituted C2-C12 alkynyl group, an optionally substituted C6-C10 aryl group or a 5- to 10-membered heteroaryl group; or R5 and R6 together with the nitrogen atom to which they are attached form a ring, and G5, R e and R f are as defined in claim 1.

11. The compound according to claim 3, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, characterized in that, The compound has the structure shown in formula (I-B-1): wherein m and n are each independently 1 or 3, R e and R f are as defined in claim 1.

12. The compound according to any one of claims 1 to 12, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, wherein R e and / or R f has one of the following structures:

13. The compound according to any one of claims 1 to 12, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, which is the following compound, 14. A composition comprising: a therapeutic or prophylactic agent; and a carrier for delivering the therapeutic or prophylactic agent, wherein, The carrier comprises an ionizable lipid, and the ionizable lipid comprises one or more of the compounds of formula (I) as defined in any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof.

15. The composition according to claim 14, wherein the therapeutic or prophylactic agent is selected from one or more of nucleic acid molecules, small molecule compounds, polypeptides or proteins. Preferably, the nucleic acid molecule is selected from one or more of single-stranded DNA, double-stranded DNA, short isoforms, agomir, antagomir, antisense molecules, small interfering RNA (siRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), Dicer substrate RNA (dsRNA), short hairpin RNA (shRNA), transfer RNA (tRNA), messenger RNA (mRNA), locked nucleic acid (LNA), peptide nucleic acid (PNA) or morpholino oligonucleotides.

16. The composition according to claim 15, wherein the therapeutic or prophylactic agent comprises at least one mRNA encoding an antigen or a fragment or epitope thereof. Preferably, the mRNA is monocistronic mRNA or polycistronic mRNA.

17. The composition according to claim 16, wherein the antigen is a pathogenic antigen.

18. The composition according to claim 15, wherein the mRNA comprises one or more functional nucleotide analogs selected from one or more of pseudouridine, 1-methyl-pseudouridine or 5-methylcytosine.

19. The composition according to claim 15, wherein the small molecule compound is selected from one or more of anti-tumor drugs, anti-infective drugs, local anesthetics, antidepressants, anti-convulsants, antibiotics / antibacterials, anti-fungal drugs, anti-parasitic drugs, hormones, hormone antagonists, immunomodulators, neurotransmitter antagonists, anti-glaucoma agents, anesthetics or imaging agents.

20. The composition according to claim 14, wherein the mass ratio of the carrier to the therapeutic or prophylactic agent is 5:1 to 50:

1.

21. The composition according to claim 14, wherein the composition is a nanoparticle formulation, and the average size of the nanoparticle formulation is 10 to 500 nm; or the pKa of the nanoparticle formulation is 4.5 to 8.

5.

22. The composition according to any one of claims 14 - 21, wherein, The carrier further comprises one or more neutral lipids. Preferably, the neutral lipid is selected from one or more of phosphatidylcholine, phosphatidylethanolamine, sphingomyelin, ceramide, sterols and their derivatives.

23. The composition according to any one of claims 14-21, wherein the molar ratio of the ionizable lipid to the neutral lipid is 100:1 to 5:

1.

24. The composition according to any one of claims 14-21, wherein, The carrier further comprises a steroid. Preferably, the steroid is selected from one or more of cholesterol, non-sterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatine, tomatidine, ursolic acid, α-tocopherol, corticosteroids.

25. The composition according to any one of claims 14-21, wherein the molar ratio of the ionizable lipid to the steroid is 2:1 to 4:

1.

26. The composition according to any one of claims 14-21, wherein the carrier further comprises one or more lipids capable of binding to the polymer, preferably, the lipid capable of binding to the polymer is one or more selected from PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, PEG-modified dialkylglycerol, and further preferably, the molar ratio of the ionizable lipid to the lipid capable of binding to the polymer is 100:1 to 20:

1.

27. The composition according to any one of claims 14-21, the carrier further comprises neutral lipid, structural lipid and polymer conjugate lipid, and the molar ratio of the ionizable lipid, the neutral lipid, the steroid lipid, and the polymer conjugate lipid is (15-70):(1-15):(15-35):(0-3).

28. The composition according to claim 14, wherein The composition further comprises a pharmaceutically acceptable excipient, preferably, the excipient comprises a pharmaceutically acceptable diluent.

29. Use of a compound of formula (I) according to any one of claims 1-13, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, or the composition according to any one of claims 14-28 in the preparation of a drug, preferably the drug is any one selected from gene drugs, nucleic acid vaccines, small molecule drugs, polypeptides or protein drugs.

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